Your One-Stop-Shop for Molecular Biology

Transcription

1 Your One-Stop-Shop for Molecular Biology Standard & Real-Time PCR Primers & Oligonucleotides DNA Mutagenesis & Preparation DNA Labeling Restriction & Modifying Enzymes

2 Jena Bioscience Company Profile Jena Bioscience GmbH was founded by a team of scientists from the Max-Planck-Institute for Molecular Physiology in Dortmund. years of academic know how were condensed into the company in order to develop innovative reagents and technologies for the life science market. Since the start up in 1998, the company has evolved into an established global reagent supplier with more than 3000 products on stock and a customer base in 40 countries. Jena Bioscience serves three major client groups: Research laboratories at universities, industry, government, hospitals and medical schools Pharmaceutical industry in the process from lead discovery through to pre-clinical stages Laboratory & diagnostic reagent kit producers and re-sellers Our company premises are located in the city of Jena / Germany with a subsidiary in Teltow, just in the vicinity of the German capital Berlin. Jena Bioscience s products include nucleotides and their non-natural analogs, recombinant proteins & protein production systems, reagents for the crystallization of biological macromolecules and tailor-made solutions for molecular biology and biochemistry. For the crystallization of biological macromolecules which is the bottle neck in determining the 3D-structure of any protein we offer specialized reagents for crystal screening, crystal optimization and phasing that can reduce the time for obtaining high quality crystals ready for X-ray diffraction from several years to a few days. In our chemistry division, we have hundreds of natural and modified nucleotides available on stock. In addition, with our pre-made building blocks and in-house expertise we manufacture even the most exotic nucleotide analog from mg kg scale. In the field of recombinant protein production, Jena Bioscience has developed its proprietary LEXSY technology. LEXSY (Leishmania Expression System) is based on a S1-classified unicellular organism that combines easy handling with a full eukaryotic protein folding and modification machinery including mammalian-like glycosylation. LEXSY is primarily used for the expression of proteins that are expressed at low yields or inactive in the established systems, and expression levels of 300 mg/l of culture were achieved. Our specialized reagents are complemented with a large selection of products for any molecular biology & biochemistry laboratory such as kits for Standard PCR and Real-Time PCR, oligonucleotides, cloning enzymes, mutagenesis technologies, and many more We combine highest quality standards for all our products with an individualized customer support. We establish direct lines of communication from clients to our in-house scientists, resulting in productive interactions among people with similar background and research interest who speak the same language. Furthermore, we offer support programs and attractive discount schemes for young scientists establishing their own labs. If you wish to receive more information on Jena Bioscience, just send us an to info@jenabioscience.com.

3 Table of Contents Standard PCR 4 Product Selection Guide 4 Ready-to-Use Mixes for direct gel loading 6 Ready-to-Use Mixes 6 Core Kits 7 Thermophylic Polymerases 8 Supplements 8 Lyophilisates 9 Real-Time PCR 10 qpcr Kits for Dual Labeled Fluorescent Probes 11 Dual Labeled Fluorescent Probes 12 qpcr Kits with EvaGreen 13 Deoxynucleotides (dntps) 14 Mixes / Bundles / Single Solutions / Lyophilisates 14 Oligonucleotides 15 Standard Oligos 15 Labeled Oligos 16 DNA Ladders 17 Log Scale / Linear Scale / Classic 18 DNA Sequencing 19 Sequencing Kits 19 Sequencing Service 19 Dideoxynucleotides (ddntps)/ Termination Mixes 20 DNA Mutagenesis 21 DNA Preparation and Cleanup 22 Plasmid DNA Purification / DNA Cleanup 22 Genomic DNA Purification 23 DNA Labeling 24 Fluorescent Labeling by PCR Fluorescent Labeling by Nick Translation 26 Non-fluorescent Labeling by PCR 27 Restriction Enzymes 28 Price List 28 Enzyme Finder 29 Buffer Guide 31 Quality Standard 32 Modifying Enzymes 33 3

4 Standard PCR Product Selection Guide Tailor-made solutions for a broad range of applications Product Cat.-Nr. Convenience Yield Specificity Red Load Taq Master PCR-108 Red Load Taq Master / high yield PCR-106 Red Load Hot Start Master PCR-109 Taq Master PCR-102 Taq Master / high yield PCR-101 Hot Start Master PCR-103 High Fidelity Master PCR-104 Taq Core Kit PCR-232 Taq Core Kit / high yield PCR-231 Hot Start Core Kit PCR-233 High Fidelity Core Kit PCR-234 Pfu-X Core Kit PCR-237 Taq Pol PCR-202 Taq Pol / high yield PCR-201 Hot Start Pol PCR-203 High Fidelity Pol PCR-204 Pfu-X Polymerase PCR-207 Sequencing Pol PCR-206 Red Load Taq Master Lyophilisate PCR-151 Taq Master Lyophilisate PCR-152 Hot Start Master Lyophilisate PCR-153 PCR Additives Kit PCR-2 PCR Control Kit PCR dntp PCR Mix GCamplifier PCR Gel Loading Buffer PCR Ready-to-Use Mixes / direct gel loading Ready-to-Use Mixes Core Kits Thermophilic Polymerases Ready-to-Use Lyophilisates Supplements

5 Application Standard PCR Fidelity Routine Plate PCR / optimized for minimal by-product formation based PCR and automated pipetting Routine Not PCR / optimized for high efficiency in a broad range of reaction conditions recommended for plate based PCR and automated pipetting High Plate specificity PCR / high sensitivity PCR based PCR and automated pipetting Routine Plate PCR / optimized for minimal by-product formation based PCR and automated pipetting Routine Not PCR / optimized for high efficiency in a broad range of reaction conditions recommended for plate based PCR and automated pipetting High specificity PCR / high sensitivity PCR based PCR and automated pipetting High fidelity PCR of GC-rich and other difficult templates Plate Amplification Routine Plate PCR / optimized for minimal by-product formation based PCR and automated pipetting Routine Not PCR / optimized for high efficiency in a broad range of reaction conditions recommended for plate based PCR and automated pipetting High specificity PCR / high sensitivity PCR Plate based PCR and automated pipetting High fidelity PCR of very long templates up to 30 kb, GC-rich and other difficult templates Amplification Efficient amplification with highest fidelity Pfu polymerase with higher accuracy and increased processivity Engineered Routine Plate PCR / optimized for minimal by-product formation based PCR and automated pipetting Routine PCR / optimized for high efficiency in a broad range of reaction conditions of labeled or other modified nucleotides Incorporation High specificity PCR / high sensitivity PCR based PCR and automated pipetting High fidelity PCR of very long templates up to 30 kb, GC-rich and other difficult templates Plate Amplification Efficient amplification with highest fidelity Pfu polymerase with higher accuracy and increased processivity Engineered Incorporation of ddntps and dntps at equal rates sequencing DNA Preloaded Direct tubes and plates for routine PCR, stable at room temperature loading of the PCR product onto the gel Preloaded Stable tubes and plates for routine PCR at room temperature Preloaded Stable tubes and plates for high specificity / high sensitivity PCR at room temperature Enhancer Taq for GC-rich and other difficult templates Stabilizer to increase amplification yield Quality Includes and benchmark tests / positive control of PCR reactions / internal lab standard primer and template for amplification of the β-actin gene from human DNA Amplification Used of GC-rich DNA templates instead of standard dntp mix Facilitates Available loading of DNA containing samples into wells of agarose and polyacrylamide gels in different dye combination as blue, green and orange loading buffer

6 Standard PCR Ready-to-Use Mixes for Maximum Convenience Ready-to-Use Mixes for direct gel loading contain an inherent red dye and allow the direct loading of the PCR product onto an agarose or acrylamide gel. The Taq Master mixes are recommended for use in routine PCR and ensure fast and easy preparation with a minimum of pipetting steps. The Hot Start Master is based on a heatactivatable Taq polymerase for high specificity applications. The mixes contain all reagents required for PCR (except template and primer) in a premixed 5 concentrated solution. Ready-to-Use Mixes / direct gel loading Red Load Taq Master Taq master mix for direct gel loading PCR-108S reactions Red Load Taq Master / high yield Taq master mix for direct gel loading PCR-106S reactions Red Load Hot Start Master Hot start master mix for direct gel loading PCR-109S reactions PCR-108L PCR-106L PCR-109L 45 0 reactions reactions reactions 360 Ready-to-Use Mixes are convenient premixes containing all reagents required for PCR (except template and primer) in a premixed 5 concentrated solution. Premium quality enzymes and dntps ensure the highest quality of amplification results. Ready-to-Use Mixes Taq Master Master mix of thermostable DNA polymerase PCR-102S reactions 40 PCR-102L 0 reactions 160 Taq Master / high yield Master mix of thermostable DNA polymerase PCR-101S reactions 40 PCR-101L 0 reactions 160 Hot Start Master Master mix of heat-activatable DNA polymerase for high specificity PCR-103S reactions 80 PCR-103L 0 reactions 320 High Fidelity Master Master mix of thermostable DNA polymerase for high accuracy PCR-104S reactions PCR-104L 0 reactions Taq Pol Master Mixes allow efficient amplification of long fragments PCR from Lambda phage DNA, 4 kbp fragment Taq Pol Taq Pol / high yield buffer 0 bp Ladder Taq Master Taq Master / high yield 0 bp Ladder Red Load Taq Master Red Load Taq Master / high yield

7 Standard PCR Core Kits Complete sets of reagents required for PCR Jena Bioscience Core Kits provide you with premium quality enzymes, ultrapure dntps and optimized reaction buffers. Each kit contains complete reaction buffer with MgCl ensuring superior results in a broad range of re² action conditions or template-primer combinations. The additional reaction buffer without MgCl in combination with the MgCl stock solution allows ² ² the optimization of magnesium-sensitive PCR reactions. The kit combines simple handling with high flexibility. Core Kits Taq Core Kit Kit of thermostable DNA polymerase, dntps and reaction buffer PCR-232S 200 PCR-232L 0 Taq Core Kit / high yield Kit of thermostable DNA polymerase, dntps and high yield buffer PCR-231S 200 PCR-231L 0 Hot Start Core Kit Kit of heat-activatable DNA polymerase for high specificity, dntps and hot start buffer PCR-233S 200 PCR-233L 0 High Fidelity Core Kit Kit of thermostable DNA polymerase for high accuracy, dntps and high fidelity buffer PCR-234S 56 PCR-234L Pfu-X Core Kit Kit of proofreading DNA polymerase for highest accuracy dntps and reaction buffer PCR-237S 68 PCR-237L Taq Pol shows excellent amplification at low concentrations Human EPO-gene, 560 bp fragment bp Ladder Taq Pol 0.2 u 0.4 u 0.8 u per µl reaction bp Ladder 2u High Fidelity Pol allows efficient amplification of extremely long templates Lambda DNA, 30 kbp fragment Taq Pol / high yield 0.2 u 0.4 u 0.8 u 2 u per µl reaction bp Ladder λ DNA / Hind III High Fidelity Pol 10 ng ng template template λ DNA / Hind III

8 Standard PCR Thermophilic Polymerases for Maximum Flexibility Jena Bioscience offers a broad range of optimized Thermophilic Polymerases. Make your selection from Taq polymerases for routine PCR applications, hot start polymerases for high specific amplifications or proofreading enzyme blends for high fidelity and long range PCR. All enzymes ensure reliable, high performance results and guarantee maximum success for their particular application. Thermophilic Polymerases Taq Pol Thermostable DNA polymerase PCR-202S PCR-202L 0 Taq Pol / high yield Thermostable DNA polymerase PCR-201S 200 PCR-201L 0 Hot Start Pol Heat-activatable DNA polymerase for high specificity PCR-203S 200 PCR-203L 0 High Fidelity Pol Thermostable DNA polymerase for high accuracy PCR-204S 48 PCR-204L Pfu-X Polymerase Proofreading DNA polymerase for highest accuracy PCR-207S 60 PCR-207L Sequencing Pol Taq Pol mutant for incorporation of ddntps PCR-206S PCR-206L Supplements Standard PCR Supplements are convenient tools for routine applications or optimization of difficult primer-template combinations. They are recommended to facilitate the amplification of GC-rich structures, to enhance the yield or to serve as an internal lab standard. Supplements Gel Loading Buffer with DNA Stain Loading buffer for agarose or polyacrylamide gels with EvaGreen fluorescent DNA stain PCR Gel Loading Buffer Loading buffer for agarose or polyacrylamide gels PCR-4 5 EvaGreen Fluorescent Gel Stain Staining dye for DNA gel electrophoresis PCR-6 µl 40 PCR Control Kit Amplification of a beta-actin gene fragment from human genomic DNA PCR-3 0 reactions dntp PCR Mix GCamplifier Modified dntp mix for amplification of GC-rich sequences PCR-7 µl 80 PCR Additives Kit Stabilizer and enhancer PCR-2 0 reactions 40

9 Standard PCR Lyophilisates Preloaded and Stable at Room Temperature Ready-to-Use Lyophilisates are deliverd in PCR reaction tube strips or 96-well plates preloaded with a complete master mix in a dry, room temperature stable format. The lyophilisates combine highest performance with convenience of use and stability. There is no need for freezing, thawing or pipetting on ice. The few remaining pipetting steps minimize the risk of errors or contaminations. Each vial contains polymerase, dntps and reaction buffer required for a 20 µl PCR assay. The Red Load Taq Master Lyophilisate contains additionally an inherent red dye allowing the direct loading of the PCR reaction product onto the gel. Simply fill up the vials with template DNA, primer and PCR-grade water and run the PCR in a thermocycler as usual. Red Load Taq Master Lyophilisate Lyophilized Taq master mix containing red gel loading dye Taq Master Lyophilisate Lyophilized Taq master mix Hot Start Master Lyophilisate Lyophilized hot start master mix Ready-to-Use Lyophilisates PCR-151S-8TS preloaded 8-tube strips PCR-151L-8TS PCR-151S-FTP preloaded 96-well plates (flat top, without skirt) PCR-151L-FTP PCR-151S-HSP preloaded 96-well plates (half skirt) PCR-151L-HSP PCR-152S-8TS preloaded 8-tube strips PCR-152L-8TS PCR-152S-FTP preloaded 96-well plates (flat top, without skirt) PCR-152L-FTP PCR-152S-HSP preloaded 96-well plates (half skirt) PCR-152L-HSP PCR-153S-8TS preloaded 8-tube strips PCR-153L-8TS PCR-153S-FTP preloaded 96-well plates (flat top, without skirt) PCR-153L-FTP PCR-153S-HSP preloaded 96-well plates (half skirt) PCR-153L-HSP 12 strips / 96 reactions 60 strips / 480 reactions 2 plates / 192 reactions 10 plates / 960 reactions 2 plates / 192 reactions 10 plates / 960 reactions 12 strips / 96 reactions 60 strips / 480 reactions 2 plates / 192 reactions 10 plates / 960 reactions 2 plates / 192 reactions 10 plates / 960 reactions 12 strips / 96 reactions 60 strips / 480 reactions 2 plates / 192 reactions 10 plates / 960 reactions 2 plates / 192 reactions plates / 960 reactions 720 Ready-to-Use Lyophilisates containing primers are custom made master mix lyophilisates comprising primers, polymerase, dntps and reaction buffer. Provide us with primer sequence and we deliver preloaded PCR tubes and plates ready-made for your special application. The only thing you still have to do is adding template DNA, filling up with water and starting the cycler! Ready-to-Use Lyophilisates containing primers Red Load Taq Master Lyophilisate containing custom primers Taq Master Lyophilisate containing custom primers Hot Start Master Lyophilisate containing custom primers preloaded 8-tube strips and 96-well plates please inquire at: pcr@jenabioscience.com

10 Real-Time PCR Jena Bioscience qpcr reagents provide exceptional sensitivity and accuracy qpcr with Dual Labeled Fluorescent Probes qpcr with EvaGreen Fluorescent DNA Stain A quantitative real-time PCR assay with fluorescent probes requires polymerase, dntps, the dual labeled fluorescent probe, primers and template DNA. The proximity of fluorophore and quencher prevents the reporter dye on the probe from fluorescing. A quantitative real-time PCR assay with EvaGreen requires polymerase, dntps, EvaGreen Fluorescent DNA Stain, primers and template DNA. The dye molecules are nonfluorescent by itself The dual labeled fluorescent probe and the PCR primers bind to their target sequences during the annealing step. The PCR primers bind to their target sequences during the annealing step. 3 During the PCR extension step, the polymerase extends the primer. When the polymerase reaches the probe, its 5 3 exonuclease activity cleaves the fluorophore from the probe. The fluorophore is released and becomes fluorescent. After complete extension the detected fluorescence intensity is proportional to the amount of accumulated PCR product. The next PCR amplification cycle will be run. 4 5 During the PCR extension step, the polymerase extends the primer. EvaGreen molecules bind to the amplicon. Due to there specific interaction with dsdna the bound dye molecules become highly fluorescent. After complete extension the detected fluorescence intensity is proportional to the amount of accumulated PCR product. The next PCR amplification cycle will be run.

11 Real-Time PCR qpcr Kits with Dual Labeled Fluorescent Probes Our qpcr Product Series is designed for the quantitative real-time analysis of DNA samples using DNA probe based detection. It provides powerful tools for quantification of sample DNA in a broad dynamic range of up to 6 orders of magnitude with exceptional sensitivity and precision. The Ready-to-Use Mixes contain UNG (Uracil-N-Glycosylase) and dutp instead of dttp to prevent carry-over contaminations of DNA from previous PCR reactions. An UNG treatment at the onset of thermal cycling removes uracil residues from du-containing DNA and prevents it from serving as template. qpcr Ready-to-Use Mixes with UNG qpcr Master with UNG / ROX Real-Time PCR Master Mix containing polymerase, UNG, dntps and reaction buffer with ROX reference dye PCR-302S reactions 130 PCR-302L 0 reactions 520 qpcr Master with UNG Real-Time PCR Master Mix containing polymerase, UNG, dntps and reaction buffer PCR-301S reactions 130 PCR-301L 0 reactions 520 qpcr Master with ROX Real-Time PCR Master Mix containing polymerase, dntps and reaction buffer with ROX reference dye PCR-312S reactions 110 PCR-312L 0 reactions 440 qpcr Master Real-Time PCR Master Mix containing polymerase, dntps and reaction buffer PCR-311S reactions 110 PCR-311L 0 reactions 440 qpcr Core Kit with ROX Real-Time PCR Kit containing polymerase, dntp Mix and reaction buffer with ROX reference dye PCR-332S reactions 90 PCR-332L 0 reactions 360 qpcr Core Kit Real-Time PCR Kit containing polymerase, dntp Mix and reaction buffer PCR-331S reactions 90 PCR-331L 0 reactions 360 ROX Reference Dye Inherent reference dye allowing normalization of non-pcr related signal variation PCR reactions Thermolabile UNG (Uracil N-Glycosylase) Prevention of carry-over contaminations of du-containing DNA from previous reactions PCR qpcr Control Kit Amplification of a beta-actin gene fragment from human genomic DNA PCR reactions 130 qpcr Core Kits qpcr Supplements 11

12 Real-Time PCR Dual Labeled Fluorescent Probes Dual Labeled Fluorescent Probes are the most widely used type of DNA probes providing a highly sensitive and specific method of detection. Each DNA probe consists of a bp long sequence-specific oligonucleotide carrying a fluorophore at the 5 end and a quencher at the 3 end. Its complementary sequence to one of the strands of the amplicon ensures the high specificity of the system. The cleavage of the probe during the extension step of each PCR cycle results in a detectable fluorescence increase proportional to the amount of accumulated PCR product. Dual Labeled Fluorescent Probes 12 Oligo: 5 reporter: quencher: up to 33 bp 1 FAM, TET, JOE or HEX 3 TAMRA 0.02 µmol scale 1 OD260 from µmol scale 5 OD260 from µmol scale 8 OD260 from 426 Oligo: 5 reporter: 3 quencher: up to 33 bp FAM, TET, JOE1, HEX or TAMRA Dabcyl 0.02 µmol scale 1 OD260 from µmol scale 5 OD260 from µmol scale 8 OD260 from 462 Oligo: 5 reporter: 3 quencher: up to 33 bp FAM, TET, CAL FLuor Gold 540, JOE1, HEX or CAL Fluor Orange 560 BHQ µmol scale 1 OD260 from µmol scale 5 OD260 from µmol scale 8 OD260 from 390 up to 33 bp Cy3, TAMRA, ROX1 or CAL FLuor Red 610 BHQ µmol scale 1 OD260 from µmol scale 5 OD260 from µmol scale 8 OD260 from 442 up to 33 bp Cy5, IRD700 BHQ µmol scale 1 OD260 from µmol scale 5 OD260 from µmol scale 8 OD260 from 577 Oligo: 5 reporter: 3 quencher: Oligo: 5 reporter: 3 quencher: 1 expected minimum yields: 0.02 µmol scale 1 OD260 ; 0.2 µmol scale 2.5 OD260 ; 1 µmol scale 6 OD260 Reporter Dyes Dye Excitation max [nm] Emission max [nm] 6-FAM TET CAL Fluor Gold JOE HEX CAL Fluor Orange Cy TAMRA ROX CAL Fluor Red Cy IRD Dark Quencher Quencher Quenching max [nm] Quenching Range [nm] TAMRA DABCYL BHQ BHQ BHQ TAMRA is widely used as quencher especially in combination with the reporter FAM. Please note that TAMRA is no dark quencher and contributes to an increase in background signal because of its own fluorescence emission. Black Hole dark quencher (BHQ) probes are an advanced alternative to TAMRA and ensure a higher signal-to-noise ratio. 2 Black Hole Quencher and BHQ are trademarks registered with the US Patent and Trade Office (USPTO) Registration Number 2,883,942 and the World Intellectual Property Organization (WIPO) registration number These compounds are protected under international patent protection filed with the USPTO under patent application 09 / 567,863 currently under allowance. Black Hole Quencher dyes are licensed for sale by Biosearch Technologies, Inc., Novato, California, USA, and these products are sold exclusively for research and development purposes only. These products may not be used for any human or veterinary clinical or diagnostic purposes or any commercial purpose without express permission from Biosearch. Further, these products may not be re-sold, distributed, re-labeled or re-packaged. Cy3, Cy5 and Cy5.5 are trademarks of Amersham Pharmacia Biotech Limited or its subsidiaries.

13 Real-Time PCR qpcr Kits with EvaGreen EvaGreen fluorescent DNA stain enables rapid analysis of any target DNA without additional sequence-specific DNA probes. EvaGreen is compatible with all common real-time PCR cyclers simply select the standard settings for SYBR Green or FAM! EvaGreen Fluorescent DNA Stain is a superior intercalator dye specially developed for real-time qpcr applications including high-resolution DNA melting curve analysis. EvaGreen is highly stable both thermally and hydrolytically, providing maximum convenience during routine handling. Combined with its high quantum yield and low interference with PCR it is the ideal fluorophore for real-time PCR and a superior replacement for competitor dyes. High-resolution DNA melting curve analysis Competitor Dye SYBR Green I extra peak 80 Excitation and emission spectra of EvaGreen is similar to SYBR Green / FAM. Excitation max: λex=0 nm, Emission max: λem=530 nm(evagreen bound to dsdna in PBS buffer ph 7.3) Low interference with PCR EvaGreen DNA melting curve analysis using EvaGreen and Competitor Dye with 4 different amplicons. Competitor Dye shows occasional formation of an extra melting peak. Relative Fluorescence EvaGreen EvaGreen is highly stable Relative Fluorescence Simply select the optical setting for SYBR Green or FAM Wavelength (nm) 13 Cycle number Degeneration of SYBR Green I within 3 hours at 99 C. EvaGreen shows no detectable decrease in fluorescence intensity. (Each fluorophore 1.2 µm in Tris-HCl buffer ph 9.0) PCR amplification plots using EvaGreen and Competitor Dye at two different concentrations. Competitor Dye exhibits significant PCR inhibition at 1 concentration while EvaGreen does not. qpcr Ready-to-Use Mixes with EvaGreen PCR-304S reactions qpcr GreenMaster with UNG / ROX Real-Time PCR Master Mix containing polymerase, UNG, dntps and reaction buffer PCR-304L 0 reactions with EvaGreen and ROX reference dye 130 PCR-303S reactions qpcr GreenMaster with UNG Real-Time PCR Master Mix containing polymerase, UNG, dntps and reaction buffer PCR-303L 0 reactions with EvaGreen 130 qpcr GreenMaster with ROX Real-Time PCR Master Mix containing polymerase, dntps and reaction buffer with EvaGreen and ROX reference dye PCR-314S reactions 110 PCR-314L 0 reactions 440 qpcr GreenMaster Real-Time PCR Master Mix containing polymerase, dntps and reaction buffer with EvaGreen PCR-313S reactions 110 PCR-313L 0 reactions 440 qpcr Green Core Kit with ROX Real-Time PCR Kit containing polymerase, dntp Mix and reaction buffer with EvaGreen and ROX reference dye PCR-334S reactions 90 PCR-334L 0 reactions 360 qpcr Green Core Kit Real-Time PCR Kit containing polymerase, dntp Mix and reaction buffer with EvaGreen PCR-333S reactions 90 PCR-333L 0 reactions 360 PCR reactions qpcr Green Core Kits qpcr Supplements EvaGreen Fluorescent DNA Stain Superior DNA intercalator dye for DNA analysis applications EvaGreen is a registered trademark and licensed for sale by Biotium, Inc., Hayward, CA, USA. SYBR is a registered trademark of Invitrogen Corporation, Carlsbad, California, USA.

14 Deoxynucleotides (dntps) Premium Quality dntps at prices you can t ignore! Jena Bioscience s enzymatic dntp manufacturing process and refined purification protocols ensure highest quality for deoxynucleotides. All our dntps are ultrapure ( > 99%) and quality checked by a set of PCR, RT-PCR and Klenow reactions. Mixes dntp Mix Premix of 10 mm datp, dctp, dgtp and dttp NU-6S 200 µl 18 dntp Mix including dutp Premix of 10 mm datp, dctp, dgtp and 20 mm dutp NU-6L 72 NU-1020S 200 µl 22 NU-1020L 88 dntp Bundle 4 mm (datp, dctp, dgtp, dttp) NU-5S µl NU-5L 4 dntp Bundle including dutp 4 mm (datp, dctp, dgtp, dutp) NU-9S µl NU-9L 4 datp, mm NU-1 57 dctp, mm NU-2 57 dgtp, mm NU-3 57 dttp, mm NU-4 57 ditp, mm NU-7 72 NU-8 72 datp NU mg dctp NU mg dgtp NU mg dttp NU mg Bundles Single Solutions 14 dutp, mm Lyophilisates datp dctp Superior performance in PCR reactions with long templates Increased sensitivity in Real-Time PCR applications Available from small (ml) to large (liters) quantities dgtp dttp Offered at very competitive prices For larger amounts than shown here, please inquire at: nucleotides@jenabioscience.com

15 Oligonucleotides Oligonucleotides Standard Oligos Custom DNA Primers from Jena Bioscience are synthetic oligonucleotides made to order with your specified sequence. They are well suitable for use in a variety of molecular biology or analytical/diagnostic applications ranging from simple PCR and sequencing to probes for quantitative gene detection. Scale [µmol] Standard Purification* OPC Purification* HPLC Purification per base Yield [OD260] per base Yield [OD260] per base Yield [OD260] 0.02** * Standard and OPC purification can only be ordered for oligos <45 bases. ** The 0.02 µmol scale can only be ordered for oligos <34 bases. Guaranteed yields apply for a 20mer / - 20 %. For oligos >33 bases we cannot give a yield guarantee. No extra charge for 5 and internal wobbles (degenerated bases). 3 wobbles require a setup fee of 20. For technical reasons we have to double the price per base for oligos >80 bases. Amount of DNA A yield of 1 OD 260 represents approximately 33 µg of single-stranded DNA with an equal number of the four bases. This corresponds to approximately 5 nmol ( µl / µm) of a 20-mer oligonucleotide. Storage Avoid repetitive freeze/thaw cycles and long term storage at concentrations below 20 µm. Aliquot oligonucleotides before freezing. Delivery Mode Storage Temperature Shelf Life Lyophilized -20 C 1 year Lyophilized Room temperature 2 months Solution -20 C 6 months Solution Room temperature 1 week Synthesis Report A comprehensive Synthesis Report comes along with every oligo, indicating its name and sequence, synthesis scale and yield (OD, µg, nmol), delivery mode (lyophilized or solution), molecular weight, melting temperature, GC-content, purification mode and quality control. 15

16 Oligonucleotides Labeled Oligos (for Dual Labeled Fluorescent Probes please refer to page 12) A large variety of Modified Oligonucleotides is available from Jena Bioscience. All prices include HPLC purification. Please note that the prices per modification are added to the price of the oligo synthesized in standard purification (see page 14). For other modifications, larger scales or further information please inquire. 5 Non-Fluorescent Labels 5 Fluorescent Labels 0.04 µmol 2 OD µmol 3-5 OD µmol 15 OD260 Phosphate C6 Amino C12 Amino Biotin Thiol * Digoxigenin Inosine Deoxyuridine Methylcytosin Modification 0.02 µmol 1 OD µmol 2 OD µmol 3-5 OD µmol 15 OD260 Modification 0.04 µmol 2 OD µmol 3-5 OD µmol 15 OD260 6-FAM, TET HEX, TAMRA JOE, ROX Fluorescein Cy3, Cy5 Cy µmol 1 OD µmol 1 OD260 IRD 700 / CAL Fluor Gold CAL Fluor Orange CAL Fluor Red Rhodamine ITC Rhodamine Green Texas Red Modification 3 Non-Fluorescent Labels Phosphate C7 Amino Biotin Thiol Methylcytosin ddc 3 Fluorescent Labels 0.02 µmol 1 OD µmol 2 OD µmol 3-5 OD µmol 15 OD260 Inosine Deoxyuridine FAM Dabcyl HEX BHQ TAMRA BHQ Fluorescein BHQ Cy3, Cy Modification Internal Non-Fluorescent Labels Internal Fluorescent Labels 0.02 µmol 1 OD µmol 2 OD µmol 3-5 OD µmol 15 OD260 Fluorecein / FITC TAMRA Cy Modification Modification 0.02 µmol 1 OD µmol 2 OD µmol 3-5 OD µmol 15 OD260 C2 Amino dt Biotin dt ** Methylcytosin Inosine Deoxyuridine Internal fluorescent modifications require a T to be attached. The 0.02 µmol scale can only be ordered for oligos <34 bases. Guaranteed yields apply for a 20mer / - 20 %. For oligos >33 bases we cannot give a yield guarantee. Prices for 3 modifications and internal modifications are valid up to 40 bases. * For 5 Thiol a set up fee of 110 per order will be charged. ** For internal Biotin dt a set up fee of 0 per order will be charged.

17 DNA Ladders DNA Ladders Log Scale DNA Ladders are well suited for standard gel electrophoresis applications. The ladders can be combined if small and large fragments need to be analyzed on the same gel. 17 Linear Scale DNA Ladders are designed to show virtually uniform spacing over a large fragment size range. DNA Fragment Separation on Agarose Gels an Overview DNA fragment size agarose gel concentration orange G running at approx. bromophenol blue running at approx. xylene cyanol running at approx. < 20 bp 3.6% 2 bp 40 bp 280 bp bp 0 bp 3.0% 2 bp 60 bp 0 bp bp 1 kb 2.4% 3 bp bp 900 bp 200 bp 2 kb 1.8% 5 bp 200 bp 1.8 kb 0 bp 5 kb 1.2% 10 bp 0 bp 4.5 kb > 1 kb 0.6% bp 1.2 kb 12 kb

18 DNA Ladders Jena Bioscience DNA Ladders allow sizing and concentration estimates of DNA fragments on agarose gels generated by PCR or restriction digest. The Log Scale and Linear Scale Ladders are supplied in ready-to-load format containing tracking dye. Log Scale DNA Ladders bp DNA Ladder bp, 0 µl, containing orange G M -212 lanes 60 bp DNA Ladder 0 bp, 0 µl, containing orange G M -213 lanes bp DNA Ladder 0 bp, 0 µl, containing orange G / xylene cyanol M -214 lanes bp DNA Ladder bp, 0 µl, containing bromophenol blue / xylene cyanol M -215 lanes 40 0 bp DNA Ladder 0 00 bp, 0 µl, containing bromophenol blue / xylene cyanol M -216 lanes 30 1 kb DNA Ladder 1 10 kb, 0 µl, containing bromophenol blue / xylene cyanol M -217 lanes 30 Low Range DNA Ladder 0 bp, 0 µl, containing orange G M -202 lanes 55 Mid Range DNA Ladder 3000 bp, 0 µl, containing bromophenol blue / xylene cyanol M -203 lanes 45 High Range DNA Ladder kb, 0 µl, containing bromophenol blue / xylene cyanol M -204 lanes 35 M -101S µg 10 M -101L 0 µg 40 M -102S µg 10 M -102L 0 µg 40 M -103S µg 10 M -103L 0 µg 40 M -104S µg 10 M -104L 0 µg 40 M -106S µg 10 M -106L 0 µg 40 M -107S µg 40 M -107L 0 µg 160 M -108S µg 40 M -108L 0 µg 160 Linear Scale DNA Ladders Classic DNA Ladders λdna / Hind III Digest λdna / EcoR I Digest λdna / EcoR I / Hind III Digest λdna / Sty I Digest λdna / BstE II Digest pbr322 / Hinf I Digest puc19 / BsiS I (Hpa II) Digest

19 DNA Sequencing DNA Sequencing Kits (sequencing with fluorescently labeled primers) Our DNA Cycle Sequencing Kit is designed for DNA sequencing based on the Sanger Method (dideoxy chain termination method). It provides a powerful tool to derive rapidly DNA and gene sequence information as required in a multitude of molecular biological and biotechnological applications. The performance of the kit is based on a specifically engineered Taq polymerase showing an equal capability of incorporating ddntps and dntps. This guarantees the generation of uniform and easy to read sequence band patterns at lowest background. A minimal band compression of GC-rich DNA regions is achieved by optimally balanced termination mixtures containing 7-deaza-dGTP. The reaction chemistry of the kit is optimized for automated DNA sequencers and requires fluorescently labeled primers. DNA Cycle Sequencing Kit for sequencing based on fluorescently labeled primers PCR - 401S reactions 1 PCR - 401L 0 reactions 600 Service Sequencing Service of plasmid DNA or PCR amplification products Sequencing of 0 to 600 bases per run Fast sequencing data transfer per Sequencing with standard or custom primers Purification of plasmid DNA or PCR fragments 19 For inquiries or further information please refer to:

20 DNA Sequencing Dideoxynucleotides (ddntps) and Termination Mixes Dideoxynucleotide (ddntp) Bundles ddntp Bundle 4 10 mm each (ddatp, ddctp, ddgtp, ddttp) NU -1019S 4x 200 µl 135 NU -1019L 4x Dideoxynucleotide (ddntp) Single Solutions 20 ddatp 10 mm NU -1015S 200 µl NU -1015L ddctp 10 mm NU -1016S 200 µl NU -1016L ddgtp 10 mm NU -1017S 200 µl NU -1017L ddttp 10 mm NU -1018S 200 µl NU -1018L Termination Mixes (containing 7-deaza-dGTP) for cycle sequencing Terminator A 1 µm each dntp (datp, dctp, dgtp / 7-deaza dgtp, dttp), 1.5 µm ddatp PCR Terminator C 1 µm each dntp (datp, dctp, dgtp / 7-deaza dgtp, dttp), 1.5 µm ddctp PCR Terminator G 1 µm each dntp (datp, dctp, dgtp / 7-deaza dgtp, dttp), 1.5 µm ddgtp PCR Terminator T 1 µm each dntp (datp, dctp, dgtp / 7-deaza dgtp, dttp), 1.5 µm ddttp PCR Termination Mixes (containing 7-deaza-dGTP) for non-cycle sequencing Terminator A 80 µm each dntp (datp, dctp, dgtp / 7-deaza dgtp, dttp), 8 µm ddatp PCR Terminator C 80 µm each dntp (datp, dctp, dgtp / 7-deaza dgtp, dttp), 8 µm ddctp PCR Terminator G 80 µm each dntp (datp, dctp, dgtp / 7-deaza dgtp, dttp), 8 µm ddgtp PCR Terminator T 80 µm each dntp (datp, dctp, dgtp / 7-deaza dgtp, dttp), 8 µm ddttp PCR PCR - 206S PCR - 206L 0 Sequencing Polymerase Sequencing Pol Taq Pol mutant for incorporation of ddntps 280

21 The unique Jena Bioscience product series for random mutagenesis provides you with ready-to-go kits for inserting random mutations into your gene of interest. All materials are accompanied by a streamlined documentation that maximizes success. Within three billion years of evolution, nature has produced a plethora of proteins simply by repeated cycles of random mutagenesis followed by in vivo selection for superior function of the encoded proteins. This example of natural evolution has guided researchers within the last two decades to develop strategies for in vitro permutation of proteins. Among the variety of strategies applied, three major powerful techniques have emerged. Mutagenesis by dntp Analogs The method can achieve rates of mutagenesis of up to 20 %. It is based on incorporation of mutagenic dntp analogs (8oxo-dGTP and dptp) into an amplified DNA fragment by a standard PCR. The mutagenic dntps are eliminated by a second PCR step in the presence of the four natural dntps, leaving highly mutated DNA ready for further Rate of mutagenesis as a function of the number of PCR cycles investigation. Mutagenesis by Error-Prone PCR Mutagenesis is performed by a PCR reaction under conditions (increased MgCl² concentration, additional MnCl² and unbalanced dntp ratio) that induce an increased error-rate of the DNA-polymerase. Simply run the PCR protocol provided in the manual and achieve rates of mutagenesis in the range of % in a single PCR step! Enhanced mutational rate by error-prone PCR compared to standard PCR reactions Mutagenesis by DNA Shuffling Developed by Stemmer (1994) DNA shuffling generates libraries by random fragmentation of one gene or a pool of related genes, followed by the reassembly of the fragments in a self-priming PCR reaction. The rates of mutagenesis are similar to the error-prone PCR but DNA shuffling allows the recombination of sequences from different, related genes. General types of DNA shuffling DNA Mutagenesis Kits JBS dntp-mutagenesis Kit Random Mutagenesis by dntp Analogs JBS Error-Prone Kit Random Mutagenesis by Error-Prone PCR JBS DNA-Shuffling Kit Random Mutagenesis by DNA Shuffling DNA Mutagenesis DNA Mutagenesis PP reactions 240 PP reactions 190 PP reactions

22 DNA Preparation and Cleanup Our Plasmid Mini-Prep Kit is designed for isolation of high-purity plasmid or cosmid DNA from cells for subsequent sequencing, restriction digests, or transformations. Spin column based preparation provides an easy and efficient way of DNA isolation without shearing or significant loss of product and allows elution in a small volume of low-salt buffer. It eliminates time consuming phenol-chloroform extraction and alcohol precipitation and can be used either in micro-centrifuges or on vacuum manifolds. The kit allows the extraction of up to 20 µg DNA per preparation kb 2 kb Jena Bioscience Competitor 6 2 ml taken from a ml overnight culture of recombinant E. coli strain with an 8 kbp plasmid. Aliquots 2 4 were purified with the Plasmid Mini-Prep Kit from Jena Bioscience and aliquots 5 7 were purified with a competitor s kit. Purified plasmid was loaded onto a 1% agarose gel. The PCR Purification Kit allows the work-up of PCR reactions (removal of nucleotides, primers, proteins, salts and other impurities). The preparation is based on a silica-membrane technology for binding DNA in highsalt and its elution in low-salt buffer. The kit provides a simple and efficient way to purify linear or circular DNA in the range from bp to 10 kbp and is optimized for working with DNA amounts from to 0 ng. It does not require any organic extractions or precipitation and guarantees high yields and reproducibility. The Agarose Gel Extraction Kit is designed for high-yield recovery of DNA from agarose gels with simultaneous removal of primers, nucleotides, proteins, salts, agarose, ethidium bromide, and other impurities. The preparation is based on a silica-membrane technology for binding DNA in high-salt and its elution in low-salt buffer. The kit provides a simple and efficient way to purify DNA in a size range between bp and 10 kbp. It does not require any organic extractions or precipitation and guarantees high yields and reproducibility. DNA Preparation Kits Plasmid Mini-Prep Kit PCR Purification Kit Agarose Gel Extraction Kit PP-203S preparations PP-203L 0 preparations PP-201S preparations PP-201L 0 preparations PP-202S preparations PP-202L 0 preparations

23 DNA Preparation and Cleanup Genomic DNA Purification Kits are designed for convenient and fast isolation of total DNA from a variety of sample sources including whole blood, bacteria, plant cells, fresh or frozen animal tissues and cells, or yeast. The solution based systems minimize DNA fragmentation that may be problematic in other spin-column / filtration based methods. Because phenol or chloroform is not used it is safe and does not produce any harmful waste. Genomic DNA purification from whole blood Yield of Purification Supplier A Supplier B Genomic DNA purification from bacteria Gram-neg. Bacteria Quality Test (17 kb long range PCR) Jena Bioscience Supplier A Supplier B Template Lane 1: Lane 2: Lane 3: Lane 4: Blood sample Lane 1: 1 µl Lane 2: 0,5 µl Lane 3: 0, µl Lane 4: 0,1 µl Supplier A Jena Bioscience 1 Gram- pos. Bacteria Jena Bioscience Supplier A 1 Quality Test Jena Bioscience Jena Bioscience Supplier A Lane 1: Lane 2: 200 ng ng ng ng E. coli Agrobacterium tumefacience Lane 1: Lane 2: C. glutamicum Bacillus subtilis Lane 1: Lane 2: Lane 3: Lane 4: 3 bp 1.2 kb 3 kb 10 kb 23 Genomic DNA purification from plant tissue Black Bean M Jena Supplier A Bioscience Genomic DNA purification from animal tissue and fungi Yellow Bean Supplier B M Jena Supplier A Bioscience Supplier B Yield of Purification M E. coli mouse tail Amplification Target Site wild hetero mutant QC by Multiplex PCR M wild hetero mutant M Genomic DNA Purification Kits Blood DNA Preparation Kit Genomic DNA purification from whole blood PP-205S preparations 80 PP-205L 400 preparations 245 Bacteria DNA Preparation Kit Genomic DNA purification from bacteria PP-206S preparations 80 PP-206L 400 preparations 245 Plant DNA Preparation Kit Genomic DNA purification from plant tissue PP-207S preparations 80 PP-207L 400 preparations 245 Animal and Fungi DNA Preparation Kit Genomic DNA purification from animal tissue and fungi PP-208S preparations 80 PP-208L 400 preparations 245 Yeast DNA Preparation Kit Genomic DNA purification from yeast PP-209S preparations PP-209L 400 preparations

24 DNA Labeling Jena Bioscience Labeling Kits guarantee superior results and performance. The fluorescently labeled dutp analogs provided in the kits are optimized for enzymatic incorporation into DNA by our proprietary linker technology. Outstanding stability and quantum yield of the thoroughly selected fluorophores combined with high incorporation rates of the dye-dutp analogs make them the ideal choice for all typical DNA labeling applications such as FISH, single molecule detection, microarray gene expression profiling and other nucleic acid hybridization assays. Emission color Labeling dye Absorption max. [nm] Emission max. [nm] Replacement for blue Atto DEAC: 430 / 477 nm 523 Alexa Fluor 488: 495 / 519 nm Fluorescein (FITC): 495 / 520 nm FAM: 495 / 520 nm Oregon Green 514: 6 / 526 nm Rhodamine green: 3 / 528 nm Rhodamine 123: 7 / 529 nm 576 Alexa Fluor 555: 555 / 565 nm Cy 3: 5 / 57 0 nm Alexa Fluor 546: 556 / 573 nm TAMRA: 546 / 576 nm Rhodamine Red: 560 / 580 nm Spectrum Orange: 559 / 588 nm green Atto yellow Atto orange Texas Red Rhodamine ITC: 572 / 596 nm Cy 3.5: 581 / 596 nm ROX: 576 / 601 nm Alexa Fluor 568: 578 / 603 nm orange Atto Alexa Fluor 594: 590 / 617 nm Alexa Fluor 610: 612 / 628 nm red Atto 647N Alexa Fluor 647: 6 / 665 nm Cy 5: 643 / 667 nm UV 400 IR

25 DNA Labeling Fluorescent Labeling by PCR PCR Labeling Kits are recommended for direct labeling of DNA by PCR using Taq polymerase. The kits contains all reagents (except primer and template) required for PCR labeling providing a highly efficient, easy-to-perform and rapid labeling technology. Kits for DNA Labeling by PCR Atto4 PCR Labeling Kit Blue-green fluorescent DNA labeling by PCR PP-301S-4 10 reactions 115 PP-301L-4 reactions 460 Atto488 PCR Labeling Kit Green fluorescent DNA labeling by PCR PP-301S reactions 115 PP-301L-488 reactions 460 Atto5 PCR Labeling Kit Yellow fluorescent DNA labeling by PCR PP-301S-5 10 reactions 115 PP-301L-5 reactions 460 TexasRed PCR Labeling Kit Orange fluorescent DNA labeling by PCR PP-301S-TXR 10 reactions 115 PP-301L-TXR reactions 460 Atto590 PCR Labeling Kit Orange fluorescent DNA labeling by PCR PP-301S reactions 115 PP-301L-590 reactions 460 PP-301S-647N 10 reactions 115 PP-301L-647N reactions 460 Atto647N PCR Labeling Kit Red fluorescent DNA labeling by PCR Fluorescently Labeled dutp Atto4-dUTP-PCR Blue-green fluorescently labeled aminoallyl-dutp PP-302S-4 10 µl / 1 mm 95 PP-302L-4 µl / 1 mm 380 Atto488-dUTP-PCR Green fluorescently labeled aminoallyl-dutp PP-302S µl / 1 mm 95 PP-302L-488 µl / 1 mm 380 Atto5-dUTP-PCR Yellow fluorescently labeled aminoallyl-dutp PP-302S-5 10 µl / 1 mm 95 PP-302L-5 µl / 1 mm 380 TexasRed-dUTP-PCR Orange fluorescently labeled aminoallyl-dutp PP-302S-TXR 10 µl / 1 mm 95 PP-302L-TXR µl / 1 mm 380 Atto590-dUTP-PCR Orange fluorescently labeled aminoallyl-dutp PP-302S µl / 1 mm 95 PP-302L-590 µl / 1 mm 380 PP-302S-647N 10 µl / 1 mm 95 PP-302L-647N µl / 1 mm 380 Atto647N-dUTP-PCR Red fluorescently labeled aminoallyl-dutp

26 DNA Labeling Preparation and Cleanup Fluorescent Labeling by Nick Translation NT Labeling Kits contain all reagents (except primer and template) required for direct labeling of DNA by nick translation using DNA polymerase I / DNase I. Kits for DNA Labeling by Nick Translation 26 Atto4 NT Labeling Kit Blue-green fluorescent DNA labeling by nick translation PP-305S-4 10 reactions 115 PP-305L-4 reactions 460 Atto488 NT Labeling Kit Green fluorescent DNA labeling by nick translation PP-305S reactions 115 PP-305L-488 reactions 460 Atto5 NT Labeling Kit Yellow fluorescent DNA labeling by nick translation PP-305S-5 10 reactions 115 PP-305L-5 reactions 460 Atto590 NT Labeling Kit Orange fluorescent DNA labeling by nick translation PP-305S reactions 115 PP-305L-590 reactions 460 PP-305S-647N 10 reactions 115 PP-305L-647N reactions 460 Atto647N NT Labeling Kit Red fluorescent DNA labeling by nick translation Fluorescently Labeled dutp Atto4-dUTP-NT Blue-green fluorescently labeled aminoallyl-dutp PP-306S-4 10 µl / 1 mm 95 PP-306L-4 µl / 1 mm 380 Atto488-dUTP-NT Green fluorescently labeled aminoallyl-dutp PP-306S µl / 1 mm 95 PP-306L-488 µl / 1 mm 380 Atto5-dUTP-NT Yellow fluorescently labeled aminoallyl-dutp PP-306S-5 10 µl / 1 mm 95 PP-306L-5 µl / 1 mm 380 Atto590-dUTP-NT Orange fluorescently labeled aminoallyl-dutp PP-306S µl / 1 mm 95 PP-306L-590 µl / 1 mm 380 PP-306S-647N 10 µl / 1 mm 95 PP-306L-647N µl / 1 mm 380 Atto647N-dUTP-NT Red fluorescently labeled aminoallyl-dutp Atto 488 Labeling Kit Atto 5 Labeling Kit Atto 590 Labeling Kit

27 DNA Preparation and Cleanup Non-fluorescent Labeling by PCR Labeling Kits Biotin PCR Labeling Kit Kit for non-fluorescent DNA labeling by PCR PP-303S-BIO 20 reactions 115 PP-303L-BIO reactions 460 Labeled dntps Biotin-dATP-PCR Non-fluorescently labeled propargylamino-datp PP-314S-BIO 10 µl / 1 mm 90 PP-314L-BIO µl / 1 mm 360 Biotin-dCTP-PCR Non-fluorescently labeled propagylamino-dctp PP-324S-BIO 10 µl / 1 mm 60 PP-324L-BIO µl / 1 mm 240 Biotin-dUTP-PCR Non-fluorescently labeled aminoallyl-dutp PP-304S-BIO 20 µl / 1 mm 90 PP-304L-BIO µl / 1 mm 360 Our Assay Kits, containing fluorescent streptavidin or avidin, are perfectly suited for binding and fluorescent detection of immobilized biotinylated samples such as proteins and DNA. Fluorescent streptavidin is a 60 kda tetrameric protein purified from the bacterium Streptomyces avidinii and labeled with fluorophores providing high fluorescence intensities. Each subunit binds one biotin molecule with high affinity. Streptavidin is not glycosylated and shows much less nonspecific interactions than avidin. Streptavidin Assay Kits Streptavidin-Atto4 Assay Kit FP assays 190 Streptavidin-Atto488 Assay Kit FP assays 190 Streptavidin-Atto532 Assay Kit FP assays 190 Streptavidin-Atto5 Assay Kit FP assays 190 Streptavidin-Atto590 Assay Kit FP assays 190 FP N assays 190 FP assays 190 Streptavidin-Atto647N Assay Kit Streptavidin-Atto655 Assay Kit Fluorescent avidin is a 66 kda tetrameric glycoprotein labeled with fluorophores providing high fluorescence intensities. Each subunit binds one biotin molecule with high affinity. The affinity of avidin for biotin is even stronger than of streptavidin, however, avidin is highly glycosylated and therefore causes higher background than streptavidin due to nonspecific interactions. Avidin Assay Kits Avidin-Atto4 Assay Kit FP assays 180 Avidin-Atto488 Assay Kit FP assays 180 Avidin-Atto532 Assay Kit FP assays 180 Avidin-Atto5 Assay Kit FP assays 180 Avidin-Atto590 Assay Kit FP assays 180 FP N assays 180 FP assays 180 Avidin-Atto647N Assay Kit Avidin-Atto655 Assay Kit 27

28 Restriction Enzymes Jena Bioscience offers a wide variety of Restriction Enzymes at very competitive prices. Most of them allow fast digestion in five minutes. Price List Enzyme Alu I ApaL I Asu II BamH I Bcl I ( C) Bgl I 28 Bgl II BseA I (55 C) BseB I (60 C) BseC I (55 C) BshF I BsiS I (55 C) BssA I (65 C) BstE II (60 C) CspA I Dpn I EcoR I EcoR V Hind III Hinf I Hpa I Kpn I Mbo I MspC I Cat.-No. EN-101S EN-101L EN-172S EN-172L EN-102S EN-102L EN-103S EN-103L EN-104S EN-104L EN-105S EN-105L EN-106S EN-106L EN-107S EN-107L EN-108S EN-108L EN-109S EN-109L EN-110S EN-110L EN-111S EN-111L EN-112S EN-112L EN-144S EN-144L EN-113S EN-113L EN-160S EN-160L EN-114S EN-114L EN-115S EN-115L EN-116S EN-116L EN-117S EN-117L EN-118S EN-118L EN-119S EN-119L EN-120S EN-120L EN-121S EN-121L Amount 600 3,000 2,000 10,000 3,0 17,0 7,0 37,0 2,0 12,0 2,000 10,000 1,300 6,0 6 3,0 4,0 22,0 3,0 17,0 7,000 35,000 2,200 11, ,0 1,7 8, ,000 15,000,000 3,000 15,000 7,0 37,0 2,0 12,0 7 3,7 3,0 17, ,0 1,300 6,0 Price Enzyme Nae I Nco I Nhe I Not I Nru I PspP I ( C) Pst I Pvu II Rsa I Sal I Sau3A I Sca I Sfi I ( C) SgrB I Sla I Sma I ( C) SnaB I Sph I SseB I Ssp I Sst I Sty I Taq I (65 C) Xba I Cat.-No. EN-122S EN-122L EN-123S EN-123L EN-146S EN-146L EN-124S EN-124L EN-1S EN-1L EN-126S EN-126L EN-127S EN-127L EN-128S EN-128L EN-129S EN-129L EN-130S EN-130L EN-1S EN-1L EN-131S EN-131L EN-132S EN-132L EN-133S EN-133L EN-134S EN-134L EN-135S EN-135L EN-136S EN-136L EN-137S EN-137L EN-138S EN-138L EN-139S EN-139L EN-140S EN-140L EN-141S EN-141L EN-142S EN-142L EN-143S EN-143L Amount 300 1, , , , , ,0 8,000 40,000 4,0 22,0 1,000 5,000 2,000 10, ,0 1,200 6, ,000 1,600 8,000 5,000,000 1, 5,0 3 1,7 0 1,0 1,0 7, ,000 1,600 8,000 6,000 30,000 3,0 17,0 3,0 17,0 Price

29 Enzyme Enzyme available from Jena Bioscience Cleavage Site 5 --> 3 JBS enzyme Enzyme Isoschizomer available from Jena Bioscience Cleavage Site 5 --> 3 Neoschizomer available from Jena Bioscience JBS enzyme Aat I AGG[CCT SseB I BspT104 I TT[CGAA Asu II Acc III T[CCGGA BseA I BspX I AT[CGAT BseC I Acc113 I AGT[ACT Sca I BsrF I R[CCGGY BssA I Acc65 I G[GTACC {Kpn I} BssA I R[CCGGY BssA I Afa I GT[AC Rsa I BssH I C[TCGAG Sla I Afl II C[TTAAG MspC I BssT1 I C[CWWGG Sty I Age I A[CCGGT CspA I Bst2U I CC[WGG BseB I Ajn I [CCWGG {BseB I} Bst98 I C[TTAAG MspC I Alu I AG[CT Alu I BstB I TT[CGAA Asu II Alw44 I G[TGCAC ApaL I BstE II G[GTNACC BstE II ApaL I G[TGCAC ApaL I BstEN II [GATC {Dpn I}, Mbo I, Sau3A I AsiA I A[CCGGT CspA I BstKT I GAT[C {Dpn I}, {Mbo I}, {Sau3A I} Asp718 I G[GTACC {Kpn I} BstMA I CTGCA[G Pst I AspS9 I G[GNCC PspP I BstMB I [GATC {Dpn I}, Mbo I, Sau3A I Asu II TT[CGAA Asu II BstN I CC[WGG BseB I AsuNH I G[CTAGC Nhe I BstO I CC[WGG BseB I BamH I G[GATCC BamH I BstP I G[GTNACC BstE II Ban III AT[CGAT BseC I BstSN I TAC[GTA SnaB I Bbu I GCATG[C Sph I Bsu15 I AT[CGAT BseC I Bcl I T[GATCA Bcl I BsuR I Bfr I C[TTAAG MspC I [GATC {Dpn I}, Mbo I, Sau3A I CciN I GC[GGCCGC Not I GCCNNNN[NGGC Bgl I Cfr10 I R[CCGGY BssA I BfuC I Bgl I BsuTU I GG[CC BshF I AT[CGAT BseC I Bgl II A[GATCT Bgl II Cfr13 I G[GNCC PspP I Bmt I GCTAG[C {Nhe I} Cfr42 I CCGC[GG SgrB I Bpu14 I TT[CGAA Asu II Cfr9 I C[CCGGG {Sma I} Bsa29 I AT[CGAT BseC I Cla I AT[CGAT BseC I Bse118 I R[CCGGY BssA I Csp45 I TT[CGAA Asu II BseA I T[CCGGA BseA I Csp6 I G[TAC {Rsa I} BseB I CC[WGG BseB I CspA I A[CCGGT CspA I BseC I AT[CGAT BseC I Dpn I GA[TC Dpn I, {Mbo I}, {Sau3A I} BshF I GG[CC BshF I Dpn II [GATC {Dpn I}, Mbo I, Sau3A I BshT I A[CCGGT CspA I Ecl136 II GAG[CTC {Sst I} BsiS I C[CGG BsiS I Eco105 I TAC[GTA SnaB I Bsp106 I AT[CGAT BseC I Eco130 I C[CWWGG Sty I Bsp119 I TT[CGAA Asu II Eco147 I AGG[CCT SseB I Bsp13 I T[CCGGA BseA I Eco32 I GAT[ATC EcoR V [GATC {Dpn I}, Mbo I, Sau3A I Eco91 I G[GTNACC BstE II Bsp19 I C[CATGG Nco I EcoICR I GAG[CTC {Sst I} Bsp68 I TCG[CGA Nru I EcoO65 I G[GTNACC BstE II Bsp143 I GG[CC BshF I EcoR I G[AATTC EcoR I BspD I AT[CGAT BseC I EcoR II [CCWGG {BseB I} BspE I T[CCGGA BseA I EcoR V GAT[ATC EcoR V BspT I C[TTAAG MspC I EcoT14 I C[CWWGG Sty I BspAN I Restriction Enzymes Enzyme Finder 29

30 Restriction Enzymes 30 Enzyme Cleavage Site 5 --> 3 Erh I Fba I Fun II Hae III Hap II Hind III Hinf I Hpa I Hpa II Kpn I C[CWWGG T[GATCA G[AATTC GG[CC C[CGG A[AGCTT G[ANTC GTT[AAC C[CGG GGTAC[C JBS enzyme Sty I Bcl I EcoR I BshF I BsiS I Hind III Hinf I Hpa I BsiS I Kpn I Enzyme Cleavage Site 5 --> 3 JBS enzyme PspE I G[GTNACC BstE II PspG I [CCWGG {BseB I} PspP I G[GNCC PspP I Pst I CTGCA[G Pst I Pvu II CAG[CTG Pvu II Rsa I GT[AC Rsa I Kpn2 I T[CCGGA Ksp I Sac I GAGCT[C Sst I Sac II CCGC[GG SgrB I BseA I Sal I GTCGAC Sal I CCGC[GG SgrB I Sau3A I [GATC {Dpn I}, Mbo I, Sau3A I Ksp22 I T[GATCA Bcl I Sau96 I G[GNCC PspP I KspA I GTT[AAC Hpa I Sca I AGT[ACT Sca I Kzo9 I [GATC {Dpn I}, Mbo I, Sau3A I Sfi I GGCCNNNN[NGGCC Sfi I Mbo I [GATC {Dpn I}, Mbo I, Sau3A I Sfr274 I C[TCGAG Sla I Mro I T[CCGGA BseA I Sfr303 I CCGC[GG SgrB I MroN I G[CCGGC {Nae I} Sfu I TT[CGAA Asu II CCGC[GG SgrB I C[CGG BsiS I MspC I C[TTAAG MspC I Sla I C[TCGAG Sla I Mva I CC[WGG BseB I Sma I CCC[GGG Sma I Nae I GCC[GGC Nae I SnaB I TAC[GTA SnaB I Nco I C[CATGG Nco I SpaH I GCATG[C Sph I Nde II [GATC {Dpn I}, Mbo I, Sau3A I Sph I GCATG[C Sph I NgoM IV G[CCGGC {Nae I} SseB I AGG[CCT SseB I Nhe I G[CTAGC Nhe I Ssp I AAT[ATT Ssp I Not I GC[GGCCGC Not I Sst I GAGCT[C Sst I Nru I TCG[CGA Nru I Stu I AGG[CCT SseB I Nsp V TT[CGAA Asu II Sty I C[CWWGG Sty I Pae I GCATG[C Sph I Taq I T[CGA Taq I PaeR7 I C[TCGAG Sla I Tli I C[TCGAG Sla I Pal I GG[CC BshF I Vha464 I C[TTAAG MspC I Pce I AGG[CCT SseB I Xba I T[CTAGA Xba I Pdi I GCC[GGC Nae I Xho I C[TCGAG Sla I Pho I GG[CC BshF I Xma I C[CCGGG {Sma I} PinA I A[CCGGT CspA I XmaC I C[CCGGG {Sma I} Psp124B I GAGCT[C Sst I Zho I AT[CGAT BseC I Psp6 I [CCWGG {BseB I} Zrm I AGT[ACT Sca I PspA I C[CCGGG {Sma I} Msp I SgrB I Single Letter Code: R = A or G, Y = C or T, M = A or C, K = G or T, S = C or G, W = A or T, H = A or C or T, B = C or G or T, V = A or C or G, D = A or G or T, N = A or C or G or T Isoschizomers have same recognition sequence and cutting pattern. Neoschizomers (same recognition sequence but different cutting pattern) are indicated with brackets {enzyme}.

31 Restriction Enzymes Buffer Guide Restriction Enzyme Alu I ApaL I Asu II BamH I Bcl I Bgl I Bgl II BseA I BseB I BseC I BshF I BsiS I BssA I BstE II CspA I Dpn I EcoR I EcoR V Hind III Hinf I Hpa I Kpn I Mbo I MspC I Nae I Nco I Nhe I Not I Nru I PspP I Pst I Pvu II Rsa I Sal I Sau3A I Sca I Sfi I SgrB I Sla I Sma I SnaB I Sph I SseB I Ssp I Sst I Sty I Taq I Xba I JBS Reaction Reaction Conditions1 Buffer min. Time Temp. B1 B1 1 B2* BamH I Buffer B2 C Bgl I Buffer B3 BseA I Buffer 55 C B2 60 C B3 55 C B5 BsiS I Buffer 55 C BssA I Buffer 65 C BstE II Buffer 60 C CspA I Buffer Dpn I Buffer EcoR I Buffer B2 1 B2 B3 B5 Kpn I Buffer Mbo I Buffer B4 B1 B3* B5 Not I Buffer 20 min. Nru I Buffer B2 C Pst I Buffer B2 B2 B4 B2 60 min. Sca I Buffer B2 C B1* B4 B5 C SnaB I Buffer B2 B3 B3 B1 B3 Taq I Buffer 1 65 C B2 B Enzyme activity (%) B2 B3 B B Alternative Reaction Buffers Fermentas NEB Buffer B NEBuffer 2 Buffer B NEBuffer 2 Buffer G NEBuffer 2 Buffer G NEBuffer 2 Buffer G NEBuffer 2 Buffer Tango NEBuffer 4 Buffer Tango NEBuffer 4 Buffer Ecl136 II NEBuffer 1 Buffer Tango NEBuffer 4 Buffer Tango NEBuffer EcoR I Buffer G NEBuffer 2 Buffer G NEBuffer 2 Buffer Tango NEBuffer 4 Buffer B NEBuffer 2 Buffer B NEBuffer 2 Buffer B (NEBuffer 4) Buffer Tango NEBuffer 4 Buffer G NEBuffer 2 Buffer G NEBuffer 2 Buffer G NEBuffer 2 Buffer G NEBuffer 2 Buffer G NEBuffer 2 Buffer B NEBuffer 2 Buffer Tango NEBuffer 4 Buffer Tango NEBuffer 1 Buffer G NEBuffer 2 Buffer B NEBuffer 2 Buffer G NEBuffer 2 Please keep in mind that different isoschizomers with the same specificity, supplied by different suppliers, could be of distinct origin and may vary in optimal reaction conditions or other properties. For this reason we recommend the use of original Jena Bioscience reaction buffers and assay conditions to achieve best results. Information on commercially available restriction endonucleases: Roberts, R. J., Restriction Enzyme Database, NEB Inc., REBASE 2000 and Fermentas International Inc. Recommended amount of enzyme: One µl enzyme per µg DNA substrate * Requires Triton X- for optimal activity. TX- is included into the supplied reaction buffer. All reactions were carried out in the presence of µg/ml BSA. 1 31

32 Restriction Enzymes Quality Standard All restriction reactions were carried out with one µl enzyme per µg DNA substrate in the presence of BamH I digest of lambda DNA BSA ( μg/ml). Our experience indicates that it is important to use BSA in the reaction assay in order to obtain successful digestions of DNA. The presence of BSA gives complete and reproducible cleavages for a range of DNA substrates. BSA stabilizes the enzymes when digestions are perfomed for more than one hour at, since many restriction endonucleases in reaction buffers without BSA can survive at this temperature for minutes only. Furthermore, BSA binds metal ions and other chemicals which might be present in buffers or DNA preparations, thereby inactivating restriction endonucleases. Unit Definition One unit of restriction endonuclease activity is defined as the amount of enzyme required to produce a complete digest of 1 µg of substrate DNA (or fragments) in a total reaction volume of µl in 60 minutes 1 1 µg lambda DNA 2 5 min digest without BSA 3 30 min digest without BSA 4 5 min digest with BSA under optimal assay conditions as stated for each restriction endonuclease. Determination of the volume activity of restriction endonucleases 32 Restriction endonuclease activity assays are performed by adding different enzyme dilutions to the appropriate assay buffer containing 1 µg of substrate DNA. After a 60-minutes incubation at the appropriate temperature, the digestion is stopped and the DNA samples are visualized by agarose gel/ethidium bromide electrophoresis. The most diluted enzyme solution giving a complete digest is used to calculate the activity in /µl. Quality Control The results of all quality control assays are reported on the Technical Data Sheet provided with each enzyme. Overdigestion Assay An overdigestion assay was used for qualitative determination of enzyme purity and for detection of nonspecific DNases. In the overdigestion assay, increasing amounts of each restriction endonuclease (usually 10, 20, 30, 40, ) are added to 1 µg substrate DNA. After a 20-hours incubation under the recommended assay conditions, the maximum number of giving a clear, sharp, normal banding pattern is determined by agarose gel/ethidium bromide electrophoresis. To pass the test, the enzyme must yield an unaltered banding pattern under conditions of up to 600-fold overdigestion ( x hours) as compared to a 2-fold digest. If enzyme exhibits star activity at a lower than 600-fold functional excess, the product description includes information on the functional excess at which the star activity does not occur. Assay for Nonspecific Endonucleases To assay for nonspecific endonuclease contamination, each restriction endonuclease is incubated with a supercoiled plasmid substrate lacking the recognition sequence of the restriction endonuclease. A single nonspecific nick in the RF I DNA converts it to the RF II form (nicked circle). Increasing amounts of enzyme (usually 10, 20, 30, 40, ) are added to 1 µg of RF I (supercoiled form) DNA. After a 20-hours incubation under the recommended assay conditions, the two forms are distinguished on agarose gels and the conversion from RF I to RF II is determined. Ligation and Recutting Assay A ligation assay was used to determine the functional purity of the DNA after restriction enzyme digestion. Substrate DNA is completely digested with a 10- and -fold excess of the restriction endonuclease in the appropriate assay buffer, ligated with T4 DNA Ligase and recut with the same restriction enzyme. Cut, ligated and recut DNA is analyzed by agarose gel/ethidium bromide electrophoresis. A normal banding pattern indicates intact 5 and 3 termini as well as the absence of contaminating nucleases or phosphatases. Stability All Jena Bioscience restriction endonucleases are reassayed every 4-6 months. This process allows us to ensure full enzyme activity and optimal performance in every enzyme we ship. Due to the excellent results of this testing, we have extended the expiration dates of most of our enzymes to 18 months.

33 Modifying Enzymes Modifying Enzymes DNA Polymerases EN-148S 300 EN-148L 1,0 EN-151S 200 EN-151L 1,000 PCR-1S 10, PCR-1L, HIV-1 RT Human Immunodeficiency Virus 1 Reverse Transcriptase PR µg 220 HIV-1 RT, p51 subunit Human Immunodeficiency Virus 1 Reverse Transcriptase PR µg 1 HIV-1 RT, p66 subunit Human Immunodeficiency Virus 1 Reverse Transcriptase PR µg 1 HIV-2 RT Human Immunodeficiency Virus 2 Reverse Transcriptase PR µg 220 EN-149S 15,000 EN-149L,000 EN-161S 0 EN-161L 2,0 160 PR , DNase Af, thermostable DNase from Archaeoglobus fulgidus, recombinant, E. coli EN-158S 1 mg 90 EN-158L 5 mg 360 RNase T1 from Aspergillus oryzae, recombinant, E. coli EN-154S 200 k EN-154L 1,000 k RNase TA engineered, recombinant, E. coli EN-156S 0 90 EN-156L 5, Exonuclease III E. coli, recombinant, E. coli EN-157S 30 k EN-157L 1 k 360 EN ,000 / 1, Topo I Human DNA Topoisomerase I, wild type PR µg 180 Topo I, several domains available Human DNA Topoisomerase I 2 µg 180 DNA Polymerase I, Klenow Fragment Fragment of DNA Polymerase I lacking 5 Y3 exonuclease activity DNA Polymerase I, Klenow Fragment, 3 Y 5 exo Fragment of DNA Polymerase I lacking 5 Y3 and 3 Y 5 exonuclease activity Reverse Transcriptases - M-MLV Reverse Transcriptase (RNase H ) Reverse Transcriptase Ligases T4 DNA Ligase E. coli lambda lysogen NM 989 Alkaline Phosphatases Alkaline Phosphatase, Calf Intestinal (CIP) from calf intestinal mucosa 40 Nucleotide Kinases T4 dnmp Kinase, T4 deoxy-nucleotide Monophosphate Kinase Bacteriophage T4, recombinant, E. coli Nucleases Exo / S1 Kit, Exonuclease III / S1 Nuclease Kit E. coli (ExoIII), Aspergillus oryzae (S1 Nuclease), recombinant, E. coli Topoisomerases 33

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